Chemical tools for developmental biology

发育生物学化学工具

• 批准号: 10369652
• 负责人: JAMES K CHEN
• 金额: $ 77.56万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10369652
• 关键词: Biological; Biology; Cell Proliferation; Chemicals; Complex; Congenital Abnormality; Detection; Development; Developmental Biology; Directed Molecular Evolution; Dynein ATPase; Erinaceidae; Floor; Genetic; Goals; Image; Imaging Device; Ions; Kinetics; Laboratories; Lanthanoid Series Elements; Lead; Life; Light; Medial; Metals; Methods; Modeling; Molecular; Morphogenesis; Natural regeneration; Oligonucleotides; Oncogenes; Optics; Pathway interactions; Pattern; Photochemistry; Physiological; Process; Property; Proteins; RNA; Research; Resolution; Signal Pathway; Signal Transduction; Somites; Technology; Tissues; Whole Organism; Zebrafish; base; cell behavior; chemical genetics; design; genetic manipulation; imaging modality; in vivo; in vivo imaging; insight; migration; notochord; novel; optogenetics; programs; response; small molecule inhibitor; spatiotemporal; tool; transcription factor; tumorigenesis

Tissue morphogenesis and regeneration requires dynamic, robust control of cell proliferation, differentiation, and migration. To coordinate these complex cell behaviors, developmental signaling pathways are actuated with spatiotemporal precision, and their dysregulation can lead to congenital birth defects or tumorigenesis later on in life. While developmental biologists have largely relied on genetic tools to deconstruct these processes, our laboratory has taken a different approach. Over the past five years, we have explored how chemical technologies and high-throughput biology can deepen our understanding of developmental signaling and tissue patterning. Over the past five years, we have invented caged morpholino oligonucleotides that can be activated by light or enzymatically triggered, and we have used these chemical tools to gain new insights into notochord, somite, and medial floor plate development. We established methods for the ultrasensitive imaging of lanthanide-based probes, allowing their unique photophysical properties to be fully exploited for autofluorescence-free in vivo imaging. We have also discovered novel regulators of the Hedgehog pathway, including ARHGAP36, a non-canonical GLI transcription factor activator and oncogene, and the first specific small-molecule inhibitors of cytoplasmic dyneins. We now seek to build upon these accomplishments and push the boundaries of in vivo chemical biology, focusing on the photochemistry of metal ions, synthetic compounds, and proteins. We envision that developmental biology would benefit from new optically controlled technologies that match the cellular resolution and rapid kinetics of patterning mechanisms, including both graded and switch-like responses. Imaging modalities that enable the detection of RNAs, proteins, and their activities at physiological concentrations would be equally transformative. Our research plans for the next five years include the synthesis of photoactivatable morpholinos with greater dynamic and spectral range, directed evolution of optogenetic regulators for key developmental signaling pathways, and design of lanthanide-based tools for imaging biological molecules in whole organisms. We will apply these technologies in zebrafish models, taking advantage of their optical transparency and amenability to chemical and genetic manipulations. Our long-term goal is to use these new experimental capabilities to perturb and observe in vivo biology in unprecedented ways, changing how we study and understand the molecular mechanisms that give rise to multicellular form.

组织形态发生和再生需要对细胞增殖进行动态、稳健的控制， 分化和迁移。为了协调这些复杂的细胞行为，发育信号通路 是以时空精度驱动的，它们的失调可能导致先天性出生缺陷， 在以后的生活中发生肿瘤。虽然发育生物学家在很大程度上依赖于遗传工具来解构 这些过程中，我们的实验室采取了不同的方法。在过去的五年里，我们探索了 化学技术和高通量生物学如何加深我们对发育的理解 信号传导和组织模式。在过去的五年里，我们发明了笼状吗啉寡核苷酸 可以被光激活或酶促触发，我们已经使用这些化学工具来获得新的 深入了解脊索、体节和内侧底板的发育。我们建立了方法， 镧系元素为基础的探针的超灵敏成像，使其独特的物理特性，以充分 用于无自体荧光的体内成像。我们还发现了新的刺猬调节器 途径，包括ARHGAP 36，一种非经典的GLI转录因子激活因子和致癌基因，以及第一个 细胞质动力蛋白的特异性小分子抑制剂。 我们现在寻求建立在这些成就的基础上，并推动体内化学生物学的界限， 主要研究金属离子、合成化合物和蛋白质的光化学。我们设想 发育生物学将受益于新的光学控制技术， 图案化机制的分辨率和快速动力学，包括分级和开关样响应。 成像模式，能够检测RNA，蛋白质及其在生理上的活性， 浓度将同样具有变革性。我们未来五年的研究计划包括 合成具有更大动力学和光谱范围的可光活化的吗啉代， 关键发育信号通路的光遗传学调节剂，以及基于镧系元素的工具的设计， 对整个生物体中的生物分子进行成像。我们将在斑马鱼模型中应用这些技术， 它们的光学透明性和对化学和遗传操作的顺从性的优点。我们的长期 目标是利用这些新的实验能力，以前所未有的方式干扰和观察体内生物学。 方法，改变我们如何研究和理解产生多细胞形式的分子机制。